Solid forms of an antiviral compound

ABSTRACT

Crystalline solid forms of methyl {(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate (Compound I) were prepared and characterized in the solid state: 
                         
Also provided are processes of manufacture and methods of using these crystalline forms.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.15/459,785, filed Mar. 15, 2017, now U.S. Pat. No. 9,884,873, which is acontinuation of U.S. application Ser. No. 14/733,101, filed Jun. 8,2015, now U.S. Pat. No. 9,630,972, which claims priority from U.S.Application No. 62/010,919, filed Jun. 11, 2014, all of which are herebyincorporated by reference in their entirety.

BACKGROUND

The present disclosure relates generally to crystalline solid forms ofthe compound methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate,designated herein as Compound I, processes for making these forms, andtheir therapeutic methods of use.

Hepatitis C is recognized as a chronic viral disease of the liver.Although drugs targeting the liver are in wide use and have showneffectiveness, toxicity and other side effects have limited theirusefulness. Inhibitors of hepatitis C virus (HCV) are useful to limitthe establishment and progression of infection by HCV as well as indiagnostic assays for HCV.

SUMMARY

Compound I which is known to exhibit antiviral properties can besynthesized according to the methods described in WO 2013/075029.Compound I has the formula:

The present disclosure provides crystalline forms of Compound I and itssalts, co-crystals, hydrates, and solvates. Also described herein areprocesses for making the mesophase and crystalline forms of Compound Iand methods for using them in the treatment of hepatitis C.

Thus, one embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(Compound I Form I) characterized by an X-ray powder diffractogramcomprising the following peaks: 4.8, 5.2, and 6.0° 2θ±0.2° 2θ, asdetermined on a diffractometer using Cu-Kα radiation at a wavelength of1.5406 Å.

Some embodiments provided herein relate to crystalline forms of thebis-hydrochloride salt of Compound I, methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrochloride (“Compound I bis-HCl”), having the formula:

Compound I bis-HCl may provide 5 forms further described herein:Compound I bis-HCl Form II, Compound I bis-HCl Form III, Compound Ibis-HCl Form IV, Compound I bis-HCl Form V, and Compound I bis-HCl FormVI.

Another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrochloride (Compound I bis-HCl Form II) characterized by an X-raypowder diffractogram comprising the following peaks: 6.1, 7.3, and 9.6°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.

Another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrochloride (Compound I bis-HCl Form III) characterized by anX-ray powder diffractogram comprising the following peaks: 7.2 and 7.6°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.

Another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrochloride (Compound I bis-HCl Form IV) characterized by an X-raypowder diffractogram comprising the following peaks: 7.5, 11.2, and14.5° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.

Another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrochloride (Compound I bis-HCl Form V) characterized by an X-raypowder diffractogram comprising the following peaks: 7.1, 10.6, and14.1° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.

Another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrochloride (Compound I bis-HCl Form VI) characterized by an X-raypowder diffractogram comprising the following peaks: 6.7 and 7.6°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.

Some embodiments provided herein relate to crystalline forms of aphosphate complex of Compound I,{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (“Compound I Phosphate”), having the formula:

wherein X can be between 2 to 3.5. Compound I Phosphate may appear inseveral forms as further described herein: Compound I Phosphate FormVII, Compound I Phosphate Form VIII, Compound I Phosphate Form IX,Compound I Phosphate Form X, Compound I Phosphate Form XI, Compound IPhosphate Form XII, Compound I Phosphate Form XIII, Compound I PhosphateForm XIV, and Compound I Phosphate Form XV.

Another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form VII) characterized by an X-raypowder diffractogram comprising the following peaks: 7.5, 14.6, and21.6° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.

Another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form VIII) characterized by an X-raypowder diffractogram comprising the following peaks: 4.2, 8.3, and 16.0°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.

Another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form IX) characterized by an X-raypowder diffractogram comprising the following peaks: 8.4, 16.1, and16.3° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.

Another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form X) characterized by an X-ray powderdiffractogram comprising the following peaks: 6.6, 9.5, and 10.6°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.

Yet another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XI) characterized by an X-raypowder diffractogram comprising the following peaks: 8.9, 13.1, and18.1° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.

Yet another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XII) characterized by an X-raypowder diffractogram comprising the following peaks: 3.8, 7.5, and 16.9°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.

Yet another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XIII) characterized by an X-raypowder diffractogram comprising the following peaks: 4.1, 15.9, and22.9° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.

Yet another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XIV) characterized by an X-raypowder diffractogram comprising the following peaks: 3.5 and 6.9°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.

Yet another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XV) characterized by an X-raypowder diffractogram comprising the following peaks: 4.0, 23.0, and24.2° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.

Some embodiments provided herein relate to crystalline forms of aL-tartrate complex of Compound I,{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateL-tartrate (“Compound I L-tartrate”), having the following formula:

wherein X can be about 2.5 to about 3. In some embodiments, X can beabout 2.5 or about 2.9. Compound I L-tartrate may appear in severalforms as further described herein: Compound I L-tartrate Form XVI andCompound I L-tartrate Form XVII.

Yet another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateL-tartrate (Compound I L-tartrate Form XVI) characterized by an X-raypowder diffractogram comprising the following peaks: 4.1, 8.1, and 15.6°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.

Yet another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateL-tartrate (Compound I L-tartrate Form XVII) characterized by an X-raypowder diffractogram comprising the following peaks: 8.2, 15.8, and22.6° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.

Some embodiments provided herein relate to crystalline forms of abis-hydrobromide salt of Compound I,{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrobromide (“Compound I bis-HBr”), having the formula:

Yet another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrobromide (Compound I bis-HBr Form XVIII) characterized by anX-ray powder diffractogram comprising the following peaks: 6.7, 7.6, and18.9° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.

One embodiment is a composition comprising any one of Compound I Form I;Compound I bis-HCl Forms II to VI; Compound I Phosphate Forms VII toForm XV; Compound I L-tartrate Forms XVI to XVII; and Compound I bis-HBrForm XVIII.

Additionally, the invention provides in one embodiment a method fortreating a subject having HCV. The method comprises administering to thesubject a therapeutically effective amount of any one of Compound IForms I to XVIII, as described generally above.

Another embodiment is the use of any one of Compound I Form I; CompoundI bis-HCl Forms II to VI; Compound I Phosphate Forms VII to Form XV;Compound I L-tartrate Forms XVI to XVII; and Compound I bis-HBr FormXVIII for the prophylactic or therapeutic treatment of hepatitis C or ahepatitis C associated disorder.

Still an additional embodiment is the use of any one of Compound I FormI; Compound I bis-HCl Forms II to VI; Compound I Phosphate Forms VII toForm XV; Compound I L-tartrate Forms XVI to XVII; and Compound I bis-HBrForm XVIII in the manufacture of a medicament for treating hepatitis Cor a hepatitis C associated disorder in a subject.

Some embodiments provided herein relate to crystalline forms of aD-tartrate complex of Compound I,{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateL-tartrate (“Compound I D-tartrate”), having the following formula:

wherein X is in the range of from about 2 to about 4. In someembodiments X is about 2, 2.5, 3.0, 3.5, or 4.0. In other particularembodiments X is about 2.5, 3.0, or 3.5. Compound I D-tartrate mayappear in several forms as further described herein: Compound ID-tartrate Form I and Compound I D-tartrate Form II.

Yet another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateD-tartrate (Compound I D-tartrate Form I) characterized by an X-raypowder diffractogram comprising the following peaks: 4.2, 8.0, and 15.3°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.

Yet another embodiment is crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateD-tartrate (Compound I D-tartrate Form II) characterized by an X-raypowder diffractogram comprising the following peaks: 4.2, 8.2, and 15.9°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a X-ray powder diffraction (XRPD) of Compound I Form I.

FIG. 2 shows a differential scanning calorimeter (DSC) curve of CompoundI Form I.

FIG. 3 shows a thermogravimetric analysis (TGA) of Compound I Form I.

FIG. 4 shows a X-ray powder diffraction (XRPD) of Compound I bis-HClForm II.

FIG. 5 shows a differential scanning calorimeter (DSC) curve of CompoundI bis-HCl Form II.

FIG. 6 shows a thermogravimetric analysis (TGA) of Compound I bis-HClForm II.

FIG. 7 shows a X-ray powder diffraction (XRPD) of Compound I bis-HClForm III.

FIG. 8 shows a differential scanning calorimeter (DSC) curve of CompoundI bis-HCl Form III.

FIG. 9 shows a thermogravimetric analysis (TGA) of Compound I bis-HClForm III.

FIG. 10 shows a X-ray powder diffraction (XRPD) of Compound I bis-HClForm IV.

FIG. 11 shows a differential scanning calorimeter (DSC) curve ofCompound I bis-HCl Form IV.

FIG. 12 shows a thermogravimetric analysis (TGA) of Compound I bis-HClForm IV.

FIG. 13 shows a X-ray powder diffraction (XRPD) of Compound I bis-HClForm V.

FIG. 14 shows a differential scanning calorimeter (DSC) curve ofCompound I bis-HCl Form V.

FIG. 15 shows a thermogravimetric analysis (TGA) of Compound I bis-HClForm V.

FIG. 16 shows a X-ray powder diffraction (XRPD) of Compound I bis-HClForm VI.

FIG. 17 shows a differential scanning calorimeter (DSC) curve ofCompound I bis-HCl Form VI.

FIG. 18 shows a thermogravimetric analysis (TGA) of Compound I bis-HClForm VI.

FIG. 19 shows a dynamic vapor sorption (DVS) curve of Compound I bis-HClForm VI.

FIG. 20 shows a X-ray powder diffraction (XRPD) of Compound I PhosphateForm VII.

FIG. 21 shows a X-ray powder diffraction (XRPD) of Compound I PhosphateForm VIII.

FIG. 22 shows a differential scanning calorimeter (DSC) curve ofCompound I Phosphate Form VIII.

FIG. 23 shows a thermogravimetric analysis (TGA) of Compound I PhosphateForm VIII.

FIG. 24 shows a X-ray powder diffraction (XRPD) of Compound I PhosphateForm IX.

FIG. 25 shows a differential scanning calorimeter (DSC) curve ofCompound I Phosphate Form IX.

FIG. 26 shows a thermogravimetric analysis (TGA) of Compound I PhosphateForm IX.

FIG. 27 shows a X-ray powder diffraction (XRPD) of Compound I PhosphateForm X.

FIG. 28 shows a X-ray powder diffraction (XRPD) of Compound I PhosphateForm XI.

FIG. 29 shows a differential scanning calorimeter (DSC) curve ofCompound I Phosphate Form XI.

FIG. 30 shows a thermogravimetric analysis (TGA) of Compound I PhosphateForm XI.

FIG. 31 shows a X-ray powder diffraction (XRPD) of Compound I PhosphateForm XII.

FIG. 32 shows a differential scanning calorimeter (DSC) curve ofCompound I Phosphate Form XII.

FIG. 33 shows a thermogravimetric analysis (TGA) of Compound I PhosphateForm XII.

FIG. 34 shows a X-ray powder diffraction (XRPD) of Compound I PhosphateForm XIII.

FIG. 35 shows a X-ray powder diffraction (XRPD) of Compound I PhosphateForm XIV.

FIG. 36 shows a thermogravimetric analysis (TGA) of Compound I PhosphateForm XIV.

FIG. 37 shows a X-ray powder diffraction (XRPD) of Compound I L-tartrateForm XVI.

FIG. 38 shows a differential scanning calorimeter (DSC) curve ofCompound I L-tartrate Form XVI.

FIG. 39 shows a thermogravimetric analysis (TGA) of Compound IL-tartrate Form XVI.

FIG. 40 shows a X-ray powder diffraction (XRPD) of Compound I L-tartrateForm XVII.

FIG. 41 shows a X-ray powder diffraction (XRPD) of Compound I bis-HBrForm XVIII.

FIG. 42 shows differential scanning calorimeter (DSC) curve of CompoundI bis-HBr Form XVIII.

FIG. 43 shows a thermogravimetric analysis (TGA) of Compound I bis-HBrForm XVIII.

FIG. 44 shows a X-ray powder diffraction (XRPD) of mesophase Compound Ibis-HCl solids from acetone. The peak at 32° 2θ is from NaCl impurity.

FIG. 45 shows a differential scanning calorimeter (DSC) curve ofmesophase Compound I bis-HCl from acetone (dried under vacuum at roomtemperature).

FIG. 46 shows a thermogravimetric analysis (TGA) of mesophase Compound Ibis-HCl from acetone (dried under vacuum at room temperature).

FIG. 47 shows a Polarized Light Microscopy (PLM) of Compound I bis-HClForm II (MeOH solvate) (top left panel); Compound I bis-HCl Form III(EtOH solvate) (bottom left panel); Compound I bis-HCl Form IV(1-propanol solvate) (top right panel); and Compound I bis-HCl Form V(EtOH solvate) (bottom right panel).

FIG. 48 shows a X-ray powder diffraction (XRPD) of Compound I PhosphateForm XV.

FIG. 49 shows a differential scanning calorimeter (DSC) curve ofCompound I Phosphate Form XV.

FIG. 50 shows an X-ray powder diffraction (XRPD) of Compound ID-tartrate Form I.

FIG. 51 shows an X-ray powder diffraction (XRPD) of Compound ID-tartrate Form II.

FIG. 52A shows a differential scanning calorimeter (DSC) curve ofCompound I D-tartrate Form II.

FIG. 52B shows a thermogravimetric analysis (TGA) of Compound ID-tartrate Form II.

FIG. 53 shows a dynamic vapor sorption (DVS) curve of Compound ID-tartrate Form II.

DETAILED DESCRIPTION

The compound methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate,designated herein as Compound I, has the formula:

The present disclosure relates to the amorphous, mesophase, andcrystalline forms of Compound I, and processes for making the mesophaseand crystalline forms. Compound I also provides a form further describedherein as “Compound I Form I.”

Additional crystalline forms of Compound I are also further describedherein. The hydrochloride salt of Compound I,{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatehydrochloride (“Compound I HCl”), has the formula:

In some embodiments, X can be 1 or 2.

The bis-hydrochloride salt of Compound I,{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrochloride (“Compound I bis-HCl”) may provide 5 forms furtherdescribed herein: Compound I bis-HCl Form II, Compound I bis-HCl FormIII, Compound I bis-HCl Form IV, Compound I bis-HCl Form V, and CompoundI bis-HCl Form VI.

The phosphate complex of Compound I,{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (“Compound I Phosphate”), has the formula:

In some embodiments, X can be between 2 to 3.5. Compound I Phosphate mayappear in several forms as further described herein: Compound IPhosphate Form VII, Compound I Phosphate Form VIII, Compound I PhosphateForm IX, Compound I Phosphate Form X, Compound I Phosphate Form XI,Compound I Phosphate Form XII, Compound I Phosphate Form XIII, CompoundI Phosphate Form XIV, and Compound I Phosphate Form XV.

The L-tartrate complex of Compound I,{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateL-tartrate (“Compound I L-tartrate”), has the following formula:

In some embodiments, X can be about 2.5. Compound I L-tartrate mayappear in several forms as further described herein: Compound IL-tartrate Form XVI and Compound I L-tartrate Form XVII.

The D-tartrate complex of Compound I,{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateD-tartrate (“Compound I D-tartrate”), has the following formula:

wherein X is in the range of from about 2 to about 4. In someembodiments X is about 2, 2.5, 3.0, 3.5, or 4.0. In other particularembodiments X is about 2.5, 3.0, or 3.5. Compound I D-tartrate mayappear in several forms as further described herein: Compound ID-tartrate Form I and Compound I D-tartrate Form II.

The hydrobromide salt of Compound I,{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatehydrobromide (“Compound I HBr”), has the following formula:

In some embodiments, X can be 1 or 2. The bis-hydrobromide salt ofCompound I,{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrobromide (“Compound I bis-HBr”), may provide Compound I bis-HBrForm XVIII as further described herein.

Definitions

As used in the present specification, the following words and phrasesare generally intended to have the meanings as set forth below, exceptto the extent that the context in which they are used indicatesotherwise.

The term “complex” refers to a formation resulting from the interactionbetween Compound I and another molecule.

The term “solvate” refers to a complex formed by combining Compound Iand a solvent.

The term “co-crystal” refers to a crystalline material formed bycombining Compound I, or any Formula disclosed herein and one or moreco-crystal formers (i.e., a molecule, ion or atom). In certaininstances, co-crystals may have improved properties as compared to theparent form (i.e., the free molecule, zwitterion, etc.) or a salt of theparent compound. Improved properties can be increased solubility,increased dissolution, increased bioavailability, increased doseresponse, decreased hygroscopicity, a crystalline form of a normallyamorphous compound, a crystalline form of a difficult to salt orunsaltable compound, decreased form diversity, more desired morphology,and the like. Methods for making and characterizing co-crystals areknown to those of skill in the art.

Any formula or structure given herein, including Compound I, is alsointended to represent unlabeled forms as well as isotopically labeledforms of the compounds. Isotopically labeled compounds have structuresdepicted by the formulae given herein except that one or more atoms arereplaced by an atom having a selected atomic mass or mass number.Examples of isotopes that can be incorporated into compounds of thedisclosure include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorous, fluorine and chlorine, such as, but not limited to ²H(deuterium, D), ³H (tritium), ¹¹, ¹³C, ¹⁴C, ¹⁵N, ¹⁸F, ³¹P, ³²P, ³⁵S,³⁶Cl and ¹²⁵I. Various isotopically labeled compounds of the presentdisclosure, for example those into which isotopes such as ³H, ¹³C and¹⁴C are incorporated. Such isotopically labeled compounds may be usefulin metabolic studies, reaction kinetic studies, detection or imagingtechniques, such as positron emission tomography (PET) or single-photonemission computed tomography (SPECT) including drug or substrate tissuedistribution assays or in radioactive treatment of patients.

The disclosure also includes Compound I in which from 1 to “n” hydrogensattached to a carbon atom is/are replaced by deuterium, in which n isthe number of hydrogens in the molecule. Such compounds exhibitincreased resistance to metabolism and are thus useful for increasingthe half-life of Compound I when administered to a mammal. See, forexample, Foster, “Deuterium Isotope Effects in Studies of DrugMetabolism”, Trends Pharmacol. Sci. 5(12):524-527 (1984). Such compoundsare synthesized by means well known in the art, for example by employingstarting materials in which one or more hydrogen atoms have beenreplaced by deuterium.

Deuterium labeled or substituted compounds of the disclosure may haveimproved DMPK (drug metabolism and pharmacokinetics) properties,relating to distribution, metabolism and excretion (ADME). Substitutionwith heavier isotopes such as deuterium may afford certain therapeuticadvantages resulting from greater metabolic stability, for exampleincreased in vivo half-life or reduced dosage requirements. An ¹⁸Flabeled compound may be useful for PET or SPECT studies. Isotopicallylabeled compounds of this disclosure and prodrugs thereof can generallybe prepared by carrying out the procedures disclosed in the schemes orin the examples and preparations described below by substituting areadily available isotopically labeled reagent for a non-isotopicallylabeled reagent. Further, substitution with heavier isotopes,particularly deuterium (i.e., ²H or D) may afford certain therapeuticadvantages resulting from greater metabolic stability, for exampleincreased in vivo half-life or reduced dosage requirements or animprovement in therapeutic index. It is understood that deuterium inthis context is regarded as a substituent in Compound I.

The concentration of such a heavier isotope, specifically deuterium, maybe defined by an isotopic enrichment factor. In the compounds of thisdisclosure any atom not specifically designated as a particular isotopeis meant to represent any stable isotope of that atom. Unless otherwisestated, when a position is designated specifically as “H” or “hydrogen”,the position is understood to have hydrogen at its natural abundanceisotopic composition. Accordingly, in the compounds of this disclosureany atom specifically designated as a deuterium (D) is meant torepresent deuterium.

The term “therapeutically effective amount” refers to an amount that issufficient to effect treatment, as defined below, when administered to amammal in need of such treatment. The therapeutically effective amountwill vary depending upon the subject being treated, the weight and ageof the subject, the severity of the disease condition, the manner ofadministration and the like, which can readily be determined by one ofordinary skill in the art.

In addition, abbreviations as used herein have respective meanings asfollows:

μL Microliter μm Micrometer 2-MeTHF 2-methyl tetrahydrofuran ACNAcetonitrile API Active pharmaceutical ingredient BE Butyl ether BNButyronitrile DCM Dichloromethane DSC Differential scanning calorimetryDVS Dynamic vapor sorption eq. equivalents EtOAc Ethyl acetate EtOHEthanol g Gram h Hour IC Ion chromatography IPA Isopropanol IPEDiisopropyl ether IPAc Isopropyl acetate KF Karl Fischer titration kVkilovolts MEK Methyl ethyl ketone MeOH Methanol MIBK Methyl iso-butylketone mA Milliamps mg Milligram min Minute mL/ml Milliliter MTBE Methyltert-butyl ether NMR Nuclear magnetic resonance PLM Polarized lightmicroscopy RT Room temperature s Second TGA Thermogravimetric analysisTHF Tetrahydrofuran XRPD X-ray powder diffractionForms of Compound I

As described generally above, the present disclosure provides mesophaseand crystalline forms of Compound I and Compound I salts/co-crystals,which are designated as Forms I to XVI.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(Compound I Form I) is characterized by an X-ray powder diffractogramcomprising the following peaks: 4.8, 5.2, and 6.0° 2θ±0.2° 2θ, asdetermined on a diffractometer using Cu-Kα radiation at a wavelength of1.5406 Å. The diffractogram comprises additional peaks at 2.9 and 3.6°2θ±0.2° 2θ. Compound I Form I is also characterized by its full X-raypowder diffractogram as substantially shown in FIG. 1.

In some embodiments, Compound I Form I is characterized by itsdifferential scanning calorimetry (DSC) curve that comprises anendotherm at about 109° C. and an endotherm at about 177° C. Compound IForm I also is characterized by its full DSC curve as substantially asshown in FIG. 2.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrochloride (Compound I bis-HCl Form II) is characterized by anX-ray powder diffractogram comprising the following peaks: 6.1, 7.3, and9.6° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiationat a wavelength of 1.5406 Å. The diffractogram comprises additionalpeaks at 3.6 and 10.9° 2θ±0.2° 2θ. Compound I bis-HCl Form II is alsocharacterized by its full X-ray powder diffractogram as substantially asshown in FIG. 4.

In some embodiments, Compound I bis-HCl Form II is characterized by itsdifferential scanning calorimetry (DSC) curve that comprises anendotherm at about 186° C. Compound I bis-HCl Form II also ischaracterized by its full DSC curve as substantially as shown in FIG. 5.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrochloride (Compound I bis-HCl Form III) is characterized by anX-ray powder diffractogram comprising the following peaks: 7.2 and 7.6°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å. The diffractogram comprises additional peaks at3.8 and 11.4° 2θ±0.2° 2θ. Compound I bis-HCl Form III is alsocharacterized by its full X-ray powder diffractogram as substantially asshown in FIG. 7.

In some embodiments, Compound I bis-HCl Form III is characterized by itsdifferential scanning calorimetry (DSC) curve that comprises anendotherm at about 189° C. Compound I bis-HCl Form III also ischaracterized by its full DSC curve as substantially as shown in FIG. 8.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrochloride (Compound I bis-HCl Form IV) is characterized by anX-ray powder diffractogram comprising the following peaks: 7.5, 11.2,and 14.5° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å. The diffractogram comprises peaksat 3.7 and 9.8° 2θ±0.2° 2θ. Compound I bis-HCl Form IV is alsocharacterized by its full X-ray powder diffractogram as substantially asshown in FIG. 10.

In some embodiments, Compound I bis-HCl Form IV is characterized by adifferential scanning calorimetry (DSC) curve that comprises anendotherm at about 193° C. Compound I bis-HCl Form IV also ischaracterized by its full DSC curve as substantially as shown in FIG.11.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrochloride (Compound I bis-HCl Form V) is characterized by anX-ray powder diffractogram comprising the following peaks: 7.1, 10.6,and 14.1° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å. The diffractogram comprises peaksat 6.3 and 12.6° 2θ±0.2° 2θ. Compound I bis-HCl Form V is alsocharacterized by its full X-ray powder diffractogram as substantially asshown in FIG. 13.

In some embodiments, Compound I bis-HCl Form V is characterized by adifferential scanning calorimetry (DSC) curve that comprises anendotherm at about 188° C. Compound I bis-HCl Form V also ischaracterized by its full DSC curve as substantially as shown in FIG.14.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrochloride (Compound I bis-HCl Form VI) is characterized by anX-ray powder diffractogram comprising the following peaks: 6.7 and 7.6°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å. The diffractogram comprises additional peaks at3.8 and 11.4° 2θ±0.2° 2θ. Compound I bis-HCl Form VI is alsocharacterized by its full X-ray powder diffractogram as substantially asshown in FIG. 16.

In some embodiments, Compound I bis-HCl Form VI is characterized by adifferential scanning calorimetry (DSC) curve that comprises anendotherm at about 205° C. Compound I bis-HCl Form VI also ischaracterized by its full DSC curve as substantially as shown in FIG.17.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form VII) characterized by an X-raypowder diffractogram comprising the following peaks: 7.5, 14.6, and21.6° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å. The diffractogram comprisesadditional peaks at 7.2 and 19.3° 2θ±0.2° 2θ. Compound I Phosphate FormVII is also characterized by its full X-ray powder diffractogram assubstantially as shown in FIG. 20.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form VIII) characterized by an X-raypowder diffractogram comprising the following peaks: 4.2, 8.3, and 16.0°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å. The diffractogram comprises additional peaks at10.1, 11.5, and 13.1° 2θ±0.2° 2θ. Compound I Phosphate Form VIII is alsocharacterized by its full X-ray powder diffractogram as substantially asshown in FIG. 21.

In some embodiments, Compound I Phosphate Form XII is characterized by adifferential scanning calorimetry (DSC) curve that comprises anendotherm at about 181° C. Compound I Phosphate Form VIII is alsocharacterized by its full DSC curve as substantially as shown in FIG.22.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form IX) is characterized by an X-raypowder diffractogram comprising the following peaks: 8.4, 16.1, and16.3° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å. The diffractogram comprisesadditional peaks at 10.2, 20.5, and 21.7° 2θ±0.2° 2θ. Compound IPhosphate Form IX is also characterized by its full X-ray powderdiffractogram as substantially as shown in FIG. 24.

In some embodiments, Compound I Phosphate Form IX is characterized by adifferential scanning calorimetry (DSC) curve that comprises anendotherm at about 177° C. and an endotherm at about 204° C. Compound IPhosphate Form IX is also characterized by its full DSC curve assubstantially as shown in FIG. 25.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form X) characterized by an X-ray powderdiffractogram comprising the following peaks: 6.6, 9.5, and 10.6°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å. The diffractogram comprises additional peaks at3.4 and 4.3° 2θ±0.2° 2θ. Compound I Phosphate Form X is alsocharacterized by its full X-ray powder diffractogram as substantially asshown in FIG. 27.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XI) is characterized by an X-raypowder diffractogram comprising the following peaks: 8.9, 13.1, and18.1° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å. The diffractogram comprisesadditional peaks at 4.0 and 17.5° 2θ±0.2° 2θ. Compound I Phosphate FormXI is also characterized by its full X-ray powder diffractogram assubstantially as shown in FIG. 28.

In some embodiments, Compound I Phosphate Form XI is also characterizedby a differential scanning calorimetry (DSC) curve that comprises anendotherm at about 172° C. and an endotherm at about 198° C. Compound IPhosphate Form XI is also characterized by its full DSC curve assubstantially as shown in FIG. 29.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XII) is characterized by an X-raypowder diffractogram comprising the following peaks: 3.8, 7.5, and 16.9°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å. The diffractogram comprises additional peaks at8.5, 10.0, and 12.4° 2θ±0.2° 2θ. Compound I Phosphate Form XII is alsocharacterized by its full X-ray powder diffractogram as substantially asshown in FIG. 31.

In some embodiments, Compound I Phosphate Form XII is also characterizedby a differential scanning calorimetry (DSC) curve that comprises anendotherm at about 205° C. and an endotherm at about 229° C. Compound IPhosphate Form XII is also characterized by its full DSC curve assubstantially as shown in FIG. 32.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XIII) 15 characterized by an X-raypowder diffractogram comprising the following peaks: 4.1, 15.9, and22.9° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å. The diffractogram comprisesadditional peaks at 7.9, 10.0, and 17.9° 2θ±0.2° 2θ. Compound IPhosphate Form XIII is also characterized by its full X-ray powderdiffractogram as substantially as shown in FIG. 34.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XIV) is characterized by an X-raypowder diffractogram comprising the following peaks: 3.5 and 6.9°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å. The diffractogram comprises additional peaks at8.3 and 12.0° 2θ±0.2° 2θ. Compound I Phosphate Form XIV is alsocharacterized by its full X-ray powder diffractogram as substantially asshown in FIG. 35.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XV) characterized by an X-raypowder diffractogram comprising the following peaks: 4.0, 23.0, and24.2° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å. The diffractogram furthercomprises peaks at 7.8 and 15.9° 2θ±0.2° 2θ. Compound I Phosphate FormXV is also characterized by its full X-ray powder diffractogram issubstantially as shown in FIG. 48.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateL-tartrate (Compound I L-tartrate Form XVI) is characterized by an X-raypowder diffractogram comprising the following peaks: 4.1, 8.1, and 15.6°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å. The diffractogram comprises additional peaks at10.0, 12.9, and 14.6° 2θ±0.2° 2θ. Compound I L-tartrate Form XVI is alsocharacterized by its full X-ray powder diffractogram as substantially asshown in FIG. 37.

In some embodiments, Compound I L-tartrate Form XVI is characterized bya differential scanning calorimetry (DSC) curve that comprises anendotherm at about 103° C., an endotherm at about 148° C., and anendotherm at about 178° C. Compound I L-tartrate Form XVI is alsocharacterized by its full DSC curve as substantially as shown in FIG.38.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateL-tartrate (Compound I L-tartrate Form XVII) is characterized by anX-ray powder diffractogram comprising the following peaks: 8.2, 15.8,and 22.6° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å. The diffractogram comprisesadditional peaks at 4.3, 10.1, and 17.9° 2θ±0.2° 2θ. Compound IL-tartrate Form XVII is also characterized by its full X-ray powderdiffractogram as substantially as shown in FIG. 40.

Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrobromide (Compound I bis-HBr Form XVIII) 15 characterized by anX-ray powder diffractogram comprising the following peaks: 6.7, 7.6, and18.9° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å. The diffractogram comprisesadditional peaks at 11.3, 15.1, and 21.8° 2θ±0.2° 2θ. Compound I bis-HBrForm XVIII is also characterized by its full X-ray powder diffractogramas substantially as shown in FIG. 41.

In some embodiments, Compound I bis-HBr Form XVIII is characterized by adifferential scanning calorimetry (DSC) curve that comprises anendotherm at about 203° C. Compound I bis-HBr Form XVIII is alsocharacterized by its full DSC curve as substantially as shown in FIG.42.

Methods of Use

The solid forms of Compound I described herein are administered fortreatment of HCV. Administration routes include, for example, thosedescribed in any patents and patent applications incorporated byreference, such as rectal, buccal, intranasal and transdermal routes, byintra-arterial injection, intravenously, intraperitoneally,parenterally, intramuscularly, subcutaneously, orally, topically, as aninhalant, or via an impregnated or coated device such as a stent, forexample, or an artery-inserted cylindrical polymer.

Oral administration can be carried out by delivering any of the CompoundI forms by capsule or enteric coated tablets, or the like.

The compounds are preferably formulated in a unit dosage form. The term“unit dosage forms” refers to physically discrete units suitable asunitary dosages for human subjects and other mammals, each unitcontaining a predetermined quantity of active material calculated toproduce the desired therapeutic effect. The compounds are generallyadministered in a pharmaceutically effective amount.

For oral administration, each dosage unit typically contains from 0.1 mgto 2 g of a compound described herein. It will be understood, however,that the amount of the compound actually administered usually will bedetermined by a physician, in the light of the relevant circumstances,including the condition to be treated, the chosen route ofadministration, the actual compound administered and its relativeactivity, the age, weight, and response of the individual patient, theseverity of the patient's symptoms, and the like.

In one embodiment, an active ingredient (i.e., Compound I, and salts andcomplexes of Compound I, as described herein) or pharmaceuticalcomposition comprising the active ingredient are effective in treatingone or more of genotype 1 HCV infected subjects, genotype 2 HCV infectedsubjects, genotype 3 HCV infected subjects, genotype 4 HCV infectedsubjects, genotype 5 HCV infected subjects, and/or genotype 6 HCVinfected subjects. In one embodiment, the active ingredient orpharmaceutical composition comprising the active ingredient areeffective in treating genotype 1 HCV infected subjects, includinggenotype 1a and/or genotype 1b. In another embodiment, the activeingredient or pharmaceutical composition comprising the activeingredient are effective in treating genotype 2 HCV infected subjects,including genotype 2a, genotype 2b, genotype 2c and/or genotype 2d. Inanother embodiment, the active ingredient or pharmaceutical compositioncomprising the active ingredient are effective in treating genotype 3HCV infected subjects, including genotype 3a, genotype 3b, genotype 3c,genotype 3d, genotype 3e and/or genotype 3f. In another embodiment, theactive ingredient or pharmaceutical composition comprising the activeingredient are effective in treating genotype 4 HCV infected subjects,including genotype 4a, genotype 4b, genotype 4c, genotype 4d, genotype4e, genotype 4f, genotype 4g, genotype 4h, genotype 4i and/or genotype4j. In another embodiment, the active ingredient or pharmaceuticalcomposition comprising the active ingredient are effective in treatinggenotype 5 HCV infected subjects, including genotype 5a. In anotherembodiment, the active ingredient or pharmaceutical compositioncomprising the active ingredient effective in treating genotype 6 HCVinfected subjects, including genotype 6a.

In some embodiments, the active ingredient or pharmaceutical compositioncomprising the active ingredient is administered, either alone or incombination with one or more therapeutic agent(s) for treating HCV (suchas a HCV NS3 protease inhibitor or an inhibitor of HCV NS5B polymerase),for about 24 weeks, for about 16 weeks, or for about 12 weeks, or less.In further embodiments, the active ingredient or pharmaceuticalcomposition comprising the active ingredient is administered, eitheralone or in combination with one or more therapeutic agent(s) fortreating HCV (such as a HCV NS3 protease inhibitor or an inhibitor ofHCV NS5B polymerase), for about 24 weeks or less, about 22 weeks orless, about 20 weeks or less, about 18 weeks or less, about 16 weeks orless, about 12 weeks or less, about 10 weeks or less, about 8 weeks orless, or about 6 weeks or less or about 4 weeks or less. The activeingredient or pharmaceutical composition comprising the activeingredient may be administered once daily, twice daily, once every otherday, two times a week, three times a week, four times a week, or fivetimes a week.

In further embodiments, a sustained virologic response is achieved atabout 4 weeks, 6 weeks, 8 weeks, 12 weeks, or 16 weeks, or at about 20weeks, or at about 24 weeks, or at about 4 months, or at about 5 months,or at about 6 months, or at about 1 year, or at about 2 years.

EXAMPLES Example 1. Stable Form Screens

Compound I can be synthesized according to the methods described in WO2013/075029 or U.S. Provisional Application No. 62/010,813 (filedconcurrently herewith and titled “Processes for Preparing AntiviralCompounds”), both of which are incorporated by reference in theirentirety. A stable form screen was performed in an attempt to obtain acrystalline form of Compound I using 10 to 20 volumes of the primarysolvent and 5 to 10 volumes of anti-solvent (if applicable).

1.1 Compound I Form I

An additional stable form screen was performed using the same procedureas described above but included a crystalline intermediate (Compound IIshown below) as seeds.

Compound II can be synthesized according to the methods described in WO2013/075029 or U.S. Provisional Application No. 62/010,813. Needle-likeparticles were formed in butyronitrile, propionitrile, MEK/toluene,MEK/IPE and 2-pentanone/toluene. XRPD patterns of the wet solids weremostly consistent with each other with minor shifting in the peaks. Thenew form is named Compound I Form I, which is believed to beisostructural channel solvates with the respective solvents. After airdrying all solids afforded amorphous XRPD patterns.

Another stable form screen was performed using carbon (Darco G-60)treated Compound I, solvents, antisolvent (diisopropyl ether (IPE)), andseeds of Compound I Form I. This screen afforded crystalline solids fromadditional solvents as summarized in Table 1. The XRPD patterns of allof these solvates are consistent with Form I. The solvates were observedto convert to amorphous solids after drying. The XRPD patterns ofCompound I were obtained in the experimental setting as follows: 45 kV,40 mA, Kα1=1.5406 Å, scan range 2-40°, step size 0.0167°, counting time:15.875 s.

TABLE 1 Stable form screen of carbon treated Compound I Solvents PLMComments Water Amorphous Slurry Water/EtOH Amorphous Sticky phasecoating ACN/IPE Birefringent Slurry of needles MeOH/IPE Solution Seedsdissolved EtOH/IPE Solution Seeds dissolved Acetone/IPE BirefringentThick slurry of needles IPA/IPE Amorphous Sticky coating MEK/IPEBirefringent Thick slurry of needles MIBK/IPE Birefringent White pasteDCM/IPE Birefringent Thick slurry of small needles THF/IPE SolutionSeeds dissolved 2-MeTHF/IPE Amorphous slurry EtOAc/IPE BirefringentThick slurry of needles IPAc/IPE Amorphous slurry Toluene AmorphousSticky coating

The crystallinity of Compound I Form I can be improved by using abutyronitrile/butyl ether (BN/BE) mixture according to the followingprocedure.

The crystallization experiment was started with 40 to 75 mg Compound Iin 1.1 to 3.0 mL of a BN/BE in a ratio of 7:4 (anhydrous solvents). Thesample was held at RT over P₂O₅ for 23 days without agitation, andcrystals formed in the solution. Afterwards, the liquid phase wasreplaced with butyl ether and the solids were obtained by centrifuge.These solids, corresponding to Compound I Form I, were used for thesubsequent step as seed.

Purified Compound I (709.8 mg) was prepared from reflux of ethanolsolution with Darco G-60 and was added to a new vial via a filter. Whilestirring, 7 mL of anhydrous butyronitrile (BN) was added. A clear orangesolution was obtained. While stirring, 4 mL of anhydrous butyl ether(BE) was added slowly. To the solution was added 7.7 mg of Compound IForm I (from previous BN:BE crystallization experiment) as seed. Thesolution became cloudy and the seeds did not dissolve. The sample wasstirred for ˜10 minutes before the agitation was stopped. The vial wascapped and placed into ajar with some P₂O₅ solids at room temperature.After 6 days, a thin layer of bright yellow precipitate was observed onthe wall and the bottom of the vial. The liquid phase was withdrawn and3 mL of anhydrous butyl ether was added. Solids were scraped down with aspatula from the vial. The suspension was heated to about 30° C. forover half hour period and was held for ˜1 hour before cooling to 20° C.at about 0.1° C./min (without agitation). The sample was stored in a jarwith P₂O₅ solids for 5 days. The sample was vacuum filtered using 0.22μm nylon filter, washed with 2×200 μL of anhydrous butyl ether, and airdried under reduced pressure for about 5 minutes.

XRPD analysis of the sample showed good very sharp peaks as shown inFIG. 1. The XRPD analysis setting was as follows: 45 kV, 40 mA,Kα1=1.5406 Å, scan range 1-40°, step size 0.0167°, counting time: 36.83s. The characteristic peaks of crystalline Compound I Form I include:2.9, 3.6, 4.8, 5.2, 6.0° 2θ (FIG. 1). The XRPD pattern of Form I wassuccessfully indexed, indicating that Form I is composed primarily of asingle crystalline phase. Extremely large unit cell volume containing upto ˜60 API molecules in the unit cell was observed. The amorphous haloobserved in the XRPD pattern could be a result of the size of the unitcell. Butyl ether stoichiometry could not be estimated. Two alternativeindexing solutions were found: monoclinic and orthorhombic.

DSC and TGA data confirmed that Form I is a solvated form. DSC shows abroad endotherm with onset at 109° C. and small endotherm with onset at177° C. (FIG. 2). TGA shows 22% weight loss below 150° C. (FIG. 3).

Example 2. Compound I Salt/Co-Crystal Screens

Salt/co-crystal screens afforded four crystalline salts or complexeswith hydrochloric acid, phosphoric acid, L-tartaric acid, andhydrobromic acid. Among those salts, Compound I bis-HCl has severalcrystalline and mesophase forms, Compound I Phosphate has eight uniqueXRPD patterns, Compound I L-tartrate has two unique XRPD patterns, andCompound I bis-HBr has one crystalline form.

2.1 Compound I bis-HCl

The mesophase solids were obtained from acetonitrile during saltformation using 10 volumes of solvent and 2 equivalents of HCl.Re-slurry of this mesophase material in acetone (10 volumes) affordedplate crystals. However, XRPD showed that although the solids fromacetone (FIG. 44) had more peaks in the XRPD pattern than those fromacetonitrile, the intensity and number of the peaks do not meet theexpectation for crystals, and it is therefore still considered amesophase. These XRPD patterns were obtained in the experimental settingas follows: 45 kV, 40 mA, Kα1=1.5406 Å, scan range 2-40°, step size0.0084°, counting time: 8.26 s. The DSC analysis of Compound I bis-HClfrom acetone was conducted using 10° C./min heating rate over the rangeof 25-400° C. and showed multiple endothermic events (FIG. 45). The TGAdata of Compound I bis-HCl from acetone were obtained using 10° C./minheating rate over the range of 25-400° C. and showed 3.6% weight lossbelow 60° C. and 6.7% weight loss at 170-220° C. (FIG. 46).

The stable form screen of Compound I bis-HCl was performed using 20solvents (5-10 volumes) in an attempt to discover all crystalline forms,as summarized in Table 2. Four crystalline solvates were discovered inthe slurries, which utilized alcohols such as methanol, ethanol and1-propanol. With ethanol, the initial precipitation was Compound Ibis-HCl Form III solids; after stirring for two weeks, the solids becameCompound I bis-HCl Form V. The shapes of these crystals appeared asneedles, shown in FIG. 47. After vacuum drying, these solvates lostsolvents easily and changed to Compound I bis-HCl Form VI. As adesolvated form, Compound I bis-HCl Form VI absorbs water quickly whenin contact with air. For example, Compound I bis-HCl Form VI contains8.8% water by KF after drying the methanol solvate and contains 5.2%water after drying the ethanol solvate. Each form was characterized byXRPD, DSC and TGA (FIGS. 4-19). TGA and DSC analysis were conductedafter the samples were dried at room temperature in vacuum oven for 1hour.

TABLE 2 Results from Stable Form Screen of Compound I bis-HCl SolubilitySolvent Initial form (mg/ml) Final form Notes Water Solution <1Amorphous EtOH/water Solution >20 Solution ACN Mesophase Mesophase GelMeOH Compound I 50 Compound Crystalline bis-HCl Form I bis-HCl II FormII EtOH Compound I 9 Compound Crystalline; bis-HCl Form I bis-HClCompound I bis-HCl Form III and III Form V Compound I bis-HCl Form Vconverted to Compound I bis-HCl Form II after vacuum drying at roomtemperature IPA Mesophase Mesophase Gel 1-Propanol Compound I 9 CompoundCrystalline; bis-HCl Form I bis-HCl Compound I bis-HCl Form IV IV FormIV converted to Compound I bis-HCl Form II after vacuum drying at roomtemperature 1-Butanol Mesophase Mesophase Gel like mesophase 2-ButanolSolution >50 Solution Acetone Mesophase 6 Mesophase Mesophase MEKAmorphous Amorphous MIBK Mesophase <1 Mesophase Mesophase DCMSolution >20 Solution Amorphous after adding heptane THF SolutionSolution Mesophase after adding heptane 2-MeTHF Amorphous AmorphousEthyl acetate Mesophase <1 Mesophase Mesophase IPAc Mesophase <1Mesophase Mesophase MTBE Amorphous <1 Amorphous Toluene Mesophase <1Mesophase Mesophase heptane Amorphous <1 Amorphous

2.1.1 Compound I bis-HCl Form II

Crystalline Compound I bis-HCl Form II was obtained from MeOH slurry (10volumes). The XRPD analysis was performed while covering the sample witha kapton film to prevent solvent evaporation. The XRPD experimentalsettings are: 45 kV, 40 mA, Kα1=1.5406 Å, scan range 2-40°, step size0.0084°, counting time: 8.26 s. The characteristic peaks of Compound Ibis-HCl Form II include: 3.6, 6.1, 7.3, 9.6, 10.9° 2θ (FIG. 4).

The DSC analysis was conducted using 2.1 mg of Compound I bis-HCl FormII sample and at a heating rate of 10° C./min over the range of 20-300°C. (FIG. 5). DSC thermogram showed multiple endothermic events includingsolvent loss below 100° C. and melting point with onset at 186° C.

The TGA data were obtained using 1.2 mg Compound I bis-HCl Form II andat a heating rate of 10° C./min over the range of 20-300° C. (FIG. 6).TGA thermogram of Compound I bis-HCl Form II showed 3.2% weight lossbelow 60° C. and 8.3% weight loss at 60-190° C. Thermal data suggestthat Compound I bis-HCl Form II is a methanol solvate.

2.1.2 Compound I bis-HCl Form III

Crystalline Compound I bis-HCl Form III was obtained from EtOH slurry(10 volumes). The following procedure was used to scale-up this form: asolution of Compound I (1.0 g) in ethanol (12.5 ml) was mixed with 37%HCl (0.28 ml) at 45° C. followed by adding Compound I bis-HCl Form IIIseeds (5 mg), which were prepared according to the method described inSection 2.1 above. The mixture was cooled from 45° C. to 20° C. Thesolids were isolated by filtration, rinsed with ethanol (2×2 ml) anddried to afford Compound I bis-HCl Form III (0.91 g).

The XRPD analysis was performed using the following experimentalsetting: 45 kV, 40 mA, Kα1=1.5406 Å, scan range 2-40°, step size0.0084°, counting time: 8.26 s. The characteristic peaks of Compound Ibis-HCl Form III include: 3.8, 7.2, 7.6, 11.4° 2θ (FIG. 7).

The DSC analysis was conducted using 2.1 mg of Compound I bis-HCl FormIII sample and a heating rate of 10° C./min over the range of 20-300° C.(FIG. 8). DSC thermogram showed multiple endothermic events includingsolvent loss below 100° C. and melting with onset at 189° C.

The TGA data were obtained using 1.2 mg Compound I bis-HCl Form III andat a heating rate of 10° C./min over the range of 20-300° C. (FIG. 9).TGA thermogram of Compound I bis-HCl Form III showed 3.6% weight lossbelow 60° C. and 7.8% weight loss at 60-190° C. Thermoanalysis datasuggest that Compound I bis-HCl Form III is an ethanol solvate.

2.1.3 Compound I bis-HCl Form IV

Crystalline Compound I bis-HCl Form IV was obtained from 1-propanolslurry (10 volumes). The XRPD analysis was performed using the followingexperimental setting: 45 kV, 40 mA, Kα1=1.5406 Å, scan range 2-40°, stepsize 0.0084°, counting time: 8.26 s. The characteristic peaks ofCompound I bis-HCl Form IV include: 3.7, 7.5, 9.8, 11.2, 14.5° 2θ (FIG.10).

The DSC analysis was conducted using 3.4 mg of Compound I bis-HCl FormIV sample and at a heating rate of 10° C./min over the range of 20-300°C. (FIG. 11). DSC thermogram showed multiple endothermic eventsincluding solvent loss below 100° C. and melting with onset at 193° C.

The TGA data were obtained using 3.5 mg Compound I bis-HCl Form IV andat a heating rate of 10° C./min over the range of 20-300° C. (FIG. 12).TGA thermogram of Compound I bis-HCl Form IV showed 5.8% weight lossbelow 60° C. and 7.7% weight loss at 60-190° C. Thermoanalysis datasuggest that Compound I bis-HCl Form IV is a 1-propanol solvate.

2.1.4 Compound I bis-HCl Form V

Crystalline Compound I bis-HCl Form V was obtained from EtOH slurry (10volumes) after two weeks. The XRPD analysis was performed using thefollowing experimental setting: 45 kV, 40 mA, Kα1=1.5406 Å, scan range2-40°, step size 0.0084°, counting time: 8.26 s. The characteristicpeaks of Compound I bis-HCl Form V include: 6.3, 7.1, 10.6, 12.6, 14.1°2θ (FIG. 13).

The DSC analysis was conducted using 2.1 mg of Compound I bis-HCl Form Vsample and at a heating rate of 10° C./min over the range of 20-300° C.(FIG. 14). DSC thermogram showed multiple endothermic events includingsolvent loss below 100° C. and melting with onset at 188° C.

The TGA data were obtained using 2.2 mg Compound I bis-HCl Form V and ata heating rate of 10° C./min over the range of 20-300° C. (FIG. 15). TGAthermogram of Compound I bis-HCl Form V showed 5.9% weight loss below60° C. and 8.7% weight loss at 60-190° C. Thermoanalysis data suggestthat Compound I bis-HCl Form V is an ethanol solvate.

2.1.5 Compound I bis-HCl Form VI

Crystalline Compound I bis-HCl Form VI was obtained after dryingsolvated Forms II through V. The XRPD analysis of Compound I bis-HClForm VI was performed using the following experimental setting: 45 kV,40 mA, Kα1=1.5406 Å, scan range 2-40°, step size 0.0167°, counting time:15.875 s. The characteristic peaks of Compound I bis-HCl Form VIinclude: 3.8, 6.7, 7.6, 11.4° 2θ (FIG. 16).

The DSC analysis was conducted using 1.4 mg of Compound I bis-HCl FormVI sample and at a heating rate of 10° C./min over the range of 20-300°C. (FIG. 17). DSC thermogram showed two broad endothermic eventsincluding solvent loss below 100° C. and melting with onset at 205° C.

The TGA data were obtained using 2.2 mg Compound I bis-HCl Form VI andat a heating rate of 10° C./min over the range of 20-300° C. (FIG. 18).TGA thermogram of Compound I bis-HCl Form VI showed 5.8% weight lossbelow 60° C. and 7.6% weight loss at 60-190° C. KF analysis showed 5.2%water. DVS analysis showed this form is hygroscopic, which explains thewater content of 5.2% (FIG. 19).

2.2 Compound I Phosphate

Crystalline Compound I Phosphate Form VII was first obtained fromEtOH/water solution with significant excess of phosphoric acid (10equivalents). Eight crystalline forms were found later (Table 3) in thestable form screening. Compound I Phosphate Form VII was observed in thewet solids from EtOH/water, and was converted to Compound I PhosphateForm VIII upon vacuum drying. Compound I Phosphate Form IX was obtainedafter air drying of Form VII. Compound I Phosphate Form X was formed byslurrying for 2 weeks either in water or in a solution of water andethanol of 0.9 water activity. This Form converted to Compound IPhosphate Form XI after drying. Compound I Phosphate Form XII wasobtained from methanol, and had the same XRPD pattern after drying.Compound I Phosphate Form XIII was formed in acetone or MEK, and itconverted to Compound I Phosphate Form VIII upon drying. Compound IPhosphate Form XIV was obtained from 9:1 acetone/water mixture or 10:12-MeTHF/water mixture, and had the same XRPD pattern upon air drying.Most of these forms were found to be hydrates except for Compound IPhosphate Form XIII, which is a channel solvate. Compound I PhosphateForm XII (wet) could be a hydrate or solvate, and converts to anisostructural hydrate after drying at ambient conditions.

TABLE 3 Crystalline forms of Compound I Phosphate Form Form Drying FormClassification Amount of Wet Form Solvent Classification conditionsafter drying after drying H₃PO₄ by IC VII Water (24 h); Hydrated Vacuumat RT; VIII Hydrate (~3 3-3.5 eq. EtOH/water (1:3) or at 45° C. eq. ofwater) Air dried IX Hydrate (~7 — eq. of water) X 0.9 water activity;Hydrated Vacuum at 45° C.; XI Hydrate (~4.4 2-2.5 eq. water (2 weeks) orair dried eq. of water) XII MeOH Possible Air dried XII Hydrate (2-3 ~3eq. Hydrate or eq. of water) solvate Note: no residual MeOH by NMR XIIIAcetone, MEK Possible Air dried VIII Hydrate (~3 — channel eq. of water)solvate XIV Acetone/water (10:1); Possible Air dried XIV Hydrate (~3~2.5 eq. 2-MeTHF/MeOH/water Hydrate eq. of water) (10:1:1);IPA/MeOH/water (10:1:1) XV EtOH Possible Air dried VIII Hydrate (~33-3.5 eq. hydrate or eq. of water) solvate

2.2.1 Compound I Phosphate Form VII

The XRPD analysis of Compound I Phosphate Form VII was performed on thewet solids obtained from EtOH/water (1:3) using the followingexperimental setting: 45 kV, 40 mA, Kα1=1.5406 Å, scan range 2-40°, stepsize 0.0167°, counting time: 15.875 s. The characteristic peaks ofCompound I Phosphate Form VII include: 7.2, 7.5, 14.6, 19.3, 21.6° 2θ(FIG. 20). Compound I Phosphate Form VII converted to Compound IPhosphate Form VIII upon vacuum drying.

2.2.2 Compound I Phosphate Form VIII

The following procedure was used to obtain Compound I Phosphate FormVIII: A solution of Compound I (1.0 g) and 85% phosphoric acid (1.3 g),water (9.0 ml) and ethanol (3.0 ml) was heated to 30° C. and seeded withCompound I Phosphate seeds (5 mg). The mixture was cooled from 30° C. to0° C. The solids were isolated by filtration, rinsed with a solution of72.5% water, 19% ethanol and 8.5% phosphoric acid (3×2 ml) and dried toafford Compound I Phosphate Form VIII (1.4 g).

The XRPD analysis of Compound I Phosphate Form VIII was performed forthe dry solids using the following experimental setting: 45 kV, 40 mA,Kα1=1.5406 Å, scan range 2-40°, step size 0.0167°, counting time: 15.875s. The characteristic peaks of Compound I Phosphate Form VIII include:4.2, 8.3, 10.1, 11.5, 13.1, 16.0° 2θ (FIG. 21).

The DSC analysis was conducted using 2-3 mg of Compound I Phosphate FormVIII sample and at a heating rate of 10° C./min over the range of20-300° C. (FIG. 22). DSC thermogram showed multiple endothermic eventsincluding solvent loss below 100° C. and the melting onset at 181° C.

The TGA data were obtained using 2-3 mg Compound I Phosphate Form VIIIand at a heating rate of 10° C./min over the range of 20-300° C. (FIG.23). TGA thermogram of Compound I Phosphate Form VIII showed 5.7% weightloss below 100° C. corresponding to the loss of residual solvent. KFanalysis showed 5.4% water content. IC analysis showed 3-3.5 equivalentsof phosphoric acid in Compound I Phosphate Form VIII.

2.2.3 Compound I Phosphate Form IX

Compound I Phosphate Form IX was obtained after drying Compound IPhosphate Form VII at ambient conditions. The XRPD analysis of CompoundI Phosphate Form IX was performed using the following experimentalsetting: 45 kV, 40 mA, Kα1=1.5406 Å, scan range 2-40°, step size0.0167°, counting time: 15.875 s. The characteristic peaks of Compound IPhosphate Form IX include: 8.4, 10.2, 16.1, 16.3, 20.5, 21.7° 2θ (FIG.24).

The DSC analysis was conducted using 2-3 mg of Compound I Phosphate FormIX sample and at a heating rate of 10° C./min over the range of 20-300°C. (FIG. 25). DSC thermogram showed multiple endothermic eventsincluding solvent loss below 150° C. and two broad endotherms withonsets at 177° C. and 204° C.

The TGA data were obtained using 5-6 mg Compound I Phosphate Form IX andat a heating rate of 10° C./min over the range of 20-300° C. (FIG. 26).TGA thermogram of Compound I Phosphate Form IX showed 9.7% weight lossbelow 120° C. corresponding to the loss of residual solvent. KF analysisshowed about 11.4% water content, corresponding to 6-7 equivalents ofwater.

2.2.4. Compound I Phosphate Form X

Compound I Phosphate Form X was obtained from 2 weeks slurry of CompoundI Phosphate Form VIII in water or EtOH/water mixture of 0.9 wateractivity (˜50 mg of Compound I Phosphate Form VIII in 1 mL of solvent).The XRPD analysis of Compound I Phosphate Form X was performed using thefollowing experimental setting: 45 kV, 40 mA, Kα1=1.5406 Å, scan range2-40°, step size 0.0167°, counting time: 15.875 s. The characteristicpeaks of Compound I Phosphate Form X include: 3.4, 4.3, 6.6, 9.5, 10.6°2θ (FIG. 27). Compound I Phosphate Form X converted to Compound IPhosphate Form XI upon air or vacuum drying. Compound I Phosphate Form Xappears to be a hydrated form.

2.2.5 Compound I Phosphate Form XI

Compound I Phosphate Form XI was obtained after drying of Compound IPhosphate Form X. The XRPD analysis of Compound I Phosphate Form XI wasperformed using the following experimental setting: 45 kV, 40 mA,Kα1=1.5406 Å, scan range 2-40°, step size 0.0167°, counting time: 15.875s. The characteristic peaks of Compound I Phosphate Form XI include:4.0, 8.9, 13.1, 17.5, 18.1° 2θ (FIG. 28).

The DSC analysis was conducted using 2-3 mg of Compound I Phosphate FormXI sample and at a heating rate of 10° C./min over the range of 20-300°C. (FIG. 29). DSC thermogram showed multiple endothermic eventsincluding solvent loss below 150° C. and two broad endotherms withonsets at 172° C. and 198° C.

The TGA data were obtained using 3-4 mg Compound I Phosphate Form XI andat a heating rate of 10° C./min over the range of 20-300° C. (FIG. 30).TGA thermogram of Compound I Phosphate Form XI showed 7.1% weight lossbelow 120° C., corresponding to the loss of residual solvent (˜4equivalents of water).

2.2.6 Compound I Phosphate Form XII

Compound I Phosphate Form XII was obtained from the slurry of Compound IPhosphate Form VIII in MeOH (˜50 mg of Compound I Phosphate Form VIII in1 mL of solvent). The XRPD analysis was performed using the followingexperimental setting: 45 kV, 40 mA, Kα1=1.5406 Å, scan range 2-40°, stepsize 0.0167°, counting time: 15.875 s. The characteristic peaks ofCompound I Phosphate Form XII include: 3.8, 7.5, 8.5, 10.0, 12.4, 16.9°2θ (FIG. 31). No significant changes were observed after drying ofCompound I Phosphate Form XII. Compound I Phosphate Form XII appears tobe a hydrated form due to 4% water content by KF and no residual MeOH byNMR.

The DSC analysis was conducted using 2-3 mg of Compound I Phosphate FormXII sample and at a heating rate of 10° C./min over the range of 20-300°C. (FIG. 32). DSC thermogram showed multiple endothermic eventsincluding solvent loss below 120° C. and two broad endotherms with onsetat 191° C. and peaks at 205° C. and 229° C., respectively.

The TGA data were obtained using 3-4 mg Compound I Phosphate Form XIIand at a heating rate of 10° C./min over the range of 20-300° C. (FIG.33). TGA thermogram of Compound I Phosphate Form XII showed 3.8% weightloss below 120° C., corresponding to the loss of residual solvent (˜2.5equivalents of water).

2.2.7 Compound I Phosphate Form XIII

Compound I Phosphate Form XIII was obtained from the slurry of CompoundI Phosphate Form VIII in acetone or MEK (˜50 mg of Compound I PhosphateForm VIII in 1 mL of solvent). The XRPD analysis was performed using thefollowing experimental setting: 45 kV, 40 mA, Kα1=1.5406 Å, scan range2-40°, step size 0.0167°, counting time: 15.875 s. The characteristicpeaks of Compound I Phosphate Form XIII include: 4.1, 7.9, 10.0, 15.9,17.9, 22.9° 2θ (FIG. 34). Compound I Phosphate Form XIII converted toCompound I Phosphate Form VIII upon air drying. Compound I PhosphateForm XIII appears to be a channel solvate.

2.2.8 Compound I Phosphate Form XIV

Compound I Phosphate Form XIV was obtained from the slurry of amorphousphosphate complex in acetone/water (10:1), in 2-MeTHF/MeOH/water(10:1:1), or in IPA/MeOH/water (10:1:1) (˜50 mg in ˜1 mL of solvent).The XRPD analysis was performed using the following experimentalsetting: 45 kV, 40 mA, Kα1=1.5406 Å, scan range 2-40°, step size0.0167°, counting time: 15.875 s. The characteristic peaks of Compound IPhosphate Form XIV include: 3.5 and 6.9° 2θ (FIG. 35). Compound IPhosphate Form XIV appears to be a hydrate based on ˜4.5% water by KF,which also contains some residual organic solvent.

The TGA data were obtained using 3-4 mg Compound I Phosphate Form XIVand at a heating rate of 10° C./min over the range of 20-300° C. (FIG.36). TGA thermogram of Compound I Phosphate Form XIV showed 4.6% weightloss below 100° C., corresponding to the loss of residual water (˜3equivalents of water), and also 3.4% weight loss at 100-150° C., mostlikely corresponding to the loss of residual solvent.

2.2.8 Compound I Phosphate Form XIV

Compound I Phosphate Form XIV was obtained using the followingprocedure: 100 mg Compound I Phosphate Form VIII was dissolved in 950 μLethanol and 50 μL 85% phosphoric acid at about 50° C. The solution wascooled and stirred at ambient temperature. After 5 days, slurry ofCompound I Phosphate Form XIV formed. The XRPD analysis was performedusing the following experimental setting: 45 kV, 40 mA, Kα1=1.5406 Å,scan range 2-40°, step size 0.0167°, counting time: 15.875 s. Thecharacteristic peaks of Compound I Phosphate Form XIV include: 4.0, 7.8,15.9, 23.0, and 24.2° 2θ (FIG. 48).

The TGA data were obtained using 2.3 mg Compound I Phosphate Form XIVand at a heating rate of 10° C./min over the range of 20-350° C. (FIG.49). TGA thermogram of Compound I Phosphate Form XIV showed about 7%weight loss below 100° C. Compound I Phosphate Form XIV appears to be asolvate.

2.3 Compound I L-Tartrate

Crystalline Compound I L-tartrate was obtained using similar conditionsas were used for the phosphate complex formation with significant excessof L-tartaric acid (10 equivalents). The dried form was designated asCompound I L-tartrate Form XVI. Compound I L-tartrate Form XVII wasobserved for the wet solids after stirring Compound I L-tartrate FormXVI in water. Compound I L-tartrate Form XVII converted to Compound IL-tartrate Form XVI upon drying.

2.3.1 Compound I L-Tartrate Form XVI

The following procedure was used to obtain Compound I L-tartrate FormXVI: a solution of Compound I (1.0 g) and L-(+)-tartaric acid (1.6 g) inethanol (10 ml) and water (1.6 ml) at 20° C. was stirred and seededCompound I L-tartrate seed crystals (5 mg). After stirring for 3 days,the solids were isolated by filtration, rinsed with ethanol (2×1 ml) anddried to afford Compound I L-tartrate Form XVI (0.47 g).

The XRPD analysis of Compound I L-tartrate Form XVI was performed usingthe following experimental setting: 45 kV, 40 mA, Kα1=1.5406 Å, scanrange 2-40°, step size 0.0167°, counting time: 15.875 s. Thecharacteristic peaks of Compound I L-tartrate Form XVI include: 4.1,8.1, 10.0, 12.9, 14.6, 15.6° 2θ (FIG. 37).

The DSC analysis was conducted using 1.4 mg of Compound I L-tartrateForm XVI sample and at a heating rate of 10° C./min over the range of20-300° C. (FIG. 38). DSC thermogram showed multiple endothermic eventsincluding solvent loss below 100° C. and endotherms with onsets at 103,148 and 178° C., respectively.

The TGA data were obtained using 2.2 mg Compound I L-tartrate Form XVIand at a heating rate of 10° C./min over the range of 20-350° C. (FIG.39). TGA thermogram of Compound I L-tartrate Form XVI showed 3.5% weightloss below 60° C. and 2.0% weight loss at 60-120° C., corresponding tothe loss of residual solvents. KF analysis showed 3.1% water content. ¹HNMR was consistent with structure of Compound I L-tartrate (with 2-3equivalents of L-tartaric acid) and showed 0.5 equivalents of residualEtOH.

2.3.2 Compound I L-Tartrate Form XVII

The XRPD analysis of Compound I L-tartrate Form XVII was performed onwet solids from water slurry using the following experimental setting:45 kV, 40 mA, Kα1=1.5406 Å, scan range 2-40°, step size 0.0167°,counting time: 15.875 s. The characteristic peaks of Compound IL-tartrate Form XVII include: 4.3, 8.2, 10.1, 15.8, 17.9, 22.6° 2θ (FIG.40). Compound I L-tartrate Form XVII converted to Compound I L-tartrateForm XVI upon air drying.

2.4 Compound I bis-HBr

Since Compound I bis-HCl was found to be crystalline, a similar attemptwas carried out to crystallize Compound I bis-HBr. The experiment wascarried out by charging 50 mg Compound I and 19 mg HBr (48% solution) in0.5 ml mixture of MeOH and MTBE. No crystals precipitated after 2 h.After seeding with Compound I HCl Form II, thick slurry was formed after1 h. XRPD shows that the pattern of precipitated crystalline solidsresembles that of Compound I HCl Form II, but they have substantialdifferences.

2.4.1 Compound I bis-HBr Form XVIII

Crystalline solids of Compound I bis-HBr Form XVIII were analyzed byXRPD, DSC, TGA, KF and IC. The XRPD analysis of Compound I bis-HBr FormXVIII was performed the following experimental setting: 45 kV, 40 mA,Kα1=1.5406 Å, scan range 2-40°, step size 0.0084°, counting time: 8.26s. The characteristic peaks of Compound I bis-HBr Form XVIII include:6.7, 7.6, 11.3, 15.1, 18.9, 21.8° 2θ (FIG. 41).

The DSC analysis was conducted using 1.9 mg of Compound I bis-HBr FormXVIII sample and at a heating rate of 10° C./min over the range of20-300° C. (FIG. 42). DSC thermogram showed multiple endothermic eventsincluding solvent loss below 100° C. and melting with onset at 203° C.

The TGA data were obtained using 2.1 mg Compound I bis-HBr Form XVIIIand at a heating rate of 10° C./min over the range of 20-300° C. (FIG.43). TGA thermogram of Compound I bis-HBr Form XVIII showed 4.2% weightloss below 60° C. and 2.7% weight loss at 60-130° C. corresponding tothe loss of residual solvents. KF analysis showed 2.5% water content. ICanalysis was performed to determine the stoichiometry of the salt andshowed about 2 equivalents of HBr.

2.5 Compound I D-Tartrate

Crystalline Compound I D-tartrate was obtained using similar conditionsas were used for Compound I L-Tartrate.

An abbreviated screen of Compound I D-tartrate was performed by stirringamorphous Compound I D-tartrate solids in 6 solvents (IPA, EtOAc,acetone, MTBE, THF and toluene) for about 2 weeks at about 21° C. Nocrystalline solids were obtained.

2.5.1 Compound I D-Tartrate Form I

The following procedure was used to obtain Compound I D-tartrate Form I:a solution of Compound I (100 mg) and D-tartaric acid (10 eq.) inethanol (0.5 mL) and water (0.5 mL) at about 21° C. was stirred forabout 2 weeks. An aliquot of the slurry was centrifuged and the wetsolids were analyzed by XRPD analysis using the following experimentalsetting: 45 kV, 40 mA, Kα1=1.5406 Å, scan range 2-40°, step size0.0084°, and counting time: 8.26 s; which showed characteristic peaksfor Compound I D-tartrate Form I at 4.2, 8.0, 9.9, 15.3, and 17.4° 2θ(FIG. 50).

2.5.2 Compound I D-Tartrate Form II

After drying Compound I D-tartrate Form I at about 40° C. under vacuumovernight, Form I became Form II resulting in a different XRPD patternwith characteristic peaks at 4.2, 8.2, 10.1, 15.9, and 17.8° 2θ (FIG.51); based on the following experimental settings: 45 kV, 40 mA,Kα1=1.5406 Å, scan range 2-40°, step size 0.084°, counting time: 8.26 s.

A DSC analysis was also conducted using 3.06 mg of Compound I D-tartrateForm II at a heating rate of 10° C./min over the range of 25-300° C.which showed endotherms with onsets at about 29, 116, and 132° C.;wherein the first two endothermic events seem to correspond to atwo-step weight loss (FIG. 52A). In addition, a TGA thermogram showedthat Compound I D-tartrate Form II had a 6.95% weight loss below 80° C.and another 1.4% weight loss between about 80 and 132° C. (FIG. 52B).Considering the significant moisture uptake in the dynamic vaporsorption (DVS) data shown in FIG. 53 between about 10-50% RH (e.g. about7%), the weight loss indicated in the TGA thermogram was most likelycaused by a loss of water.

The invention claimed is:
 1. Crystalline methyl {(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form VII) characterized by an X-raypowder diffractogram comprising the following peaks: 7.5, 14.6, and21.6° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.
 2. Compound I Phosphate Form VIIaccording to claim 1, wherein the diffractogram further comprises peaksat 7.2 and 19.3° 2θ±0.2° 2θ.
 3. Crystalline methyl {(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form X) characterized by an X-ray powderdiffractogram comprising the following peaks: 6.6, 9.5, and 10.6°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.
 4. Compound I Phosphate Form X according toclaim 3, wherein the diffractogram further comprises peaks at 3.4 and4.3° 2θ±0.2° 2θ.
 5. Crystalline methyl {(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XIII) characterized by an X-raypowder diffractogram comprising the following peaks: 4.1, 15.9, and22.9° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.
 6. Compound I Phosphate Form XIIIaccording to claim 5, wherein the diffractogram further comprises peaksat 7.9, 10.0, and 17.9° 2θ±0.2° 2θ.
 7. Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XIV) characterized by an X-raypowder diffractogram comprising the following peaks: 3.5 and 6.9°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.
 8. Compound I Phosphate Form XIV according toclaim 7, wherein the diffractogram further comprises peaks at 8.3 and12.0° 2θ±0.2° 2θ.
 9. Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatephosphate (Compound I Phosphate Form XV) characterized by an X-raypowder diffractogram comprising the following peaks: 4.0, 23.0, and24.2° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.
 10. Compound I Phosphate Form XVaccording to claim 9, wherein the diffractogram further comprises peaksat 7.8 and 15.9° 2θ±0.2° 2θ.
 11. Crystalline methyl {(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateL-tartrate (Compound I L-tartrate Form XVI) characterized by an X-raypowder diffractogram comprising the following peaks: 4.1, 8.1, and 15.6°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.
 12. Compound I L-tartrate Form XVI according toclaim 11, wherein the diffractogram further comprises peaks at 10.0,12.9, and 14.6° 2θ±0.2° 2θ.
 13. Compound I L-tartrate Form XVI accordingto claim 11, characterized by a differential scanning calorimetry (DSC)curve that comprises an endotherm at about 103° C., an endotherm atabout 148° C., and an endotherm at about 178° C.
 14. Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateL-tartrate (Compound I L-tartrate Form XVII) characterized by an X-raypowder diffractogram comprising the following peaks: 8.2, 15.8, and22.6° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.
 15. Compound I L-tartrate FormXVII according to claim 14, wherein the diffractogram further comprisespeaks at 4.3, 10.1, and 17.9° 2θ±0.2° 2θ.
 16. Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatebis-hydrobromide (Compound I bis-HBr Form XVIII) characterized by anX-ray powder diffractogram comprising the following peaks: 6.7, 7.6, and18.9° 2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kαradiation at a wavelength of 1.5406 Å.
 17. Compound I bis-HBr Form XVIIIaccording to claim 16, wherein the diffractogram further comprises peaksat 11.3, 15.1, and 21.8° 2θ±0.2° 2θ.
 18. Compound I bis-HBr Form XVIIIaccording to claim 16, characterized by a differential scanningcalorimetry (DSC) curve that comprises an endotherm at about 203° C. 19.Crystalline methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateD-tartrate (Compound I D-tartrate Form I) characterized by an X-raypowder diffractogram comprising the following peaks: 4.2, 8.0, and 15.3°2°±0.2° 2°, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.
 20. Crystalline methyl {(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2R)-2-[(methoxycarbonyl)amino]-2-phenylacetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamateD-tartrate (Compound I D-tartrate Form II) characterized by an X-raypowder diffractogram comprising the following peaks: 4.2, 8.2, and 15.9°2θ±0.2° 2θ, as determined on a diffractometer using Cu-Kα radiation at awavelength of 1.5406 Å.